EFFECT OF SOMATOSTATIN ON THE EXPRESSION OF TNF α mRNA IN MULTIORGANS OF RATS WITH ACUTE HEMORRHAGIC NECROTIC PANCREATITIS

Objectives. To study the expression of TNF α mRNA and the effecr of somatostatin on the expression of TNF α mRNA in multiorgans of rats with acute hemorrhagic necrotic panereatitis(AFINP). Methods. AHNP in the rat was induced by retrograde injection of 5% sodium tattrocholate into the bile-pancreatic duct. Somatostatin octapeptide (SS-OP) (2μg/kg)was injected into the femoral vein immediately in rats of the treatment group after inductive AHNP. Rats of the sham operative group received injection of saline. Sixty animals of the AHNP and sham operative groups at the designated time(0.2h, 0.5h, 2h, 4h, 8h, after the operation,six animals at each time point)and tweleve animals of treatment group at 4h after the operation were sacrificed for samples of pancreas, liver and lung. The expressions of TNF α mRNA within the pancreas, liver and lung were esrablished by RT-PCR. Results. TNF α mRNA became detectable in the pancreas as early as 0. 2h after inductive AHNP,while it was undetectable in other organs until 0. 5h. Expression of TNF α mRNA in each tissue inereased continuously and reached a peak at 4h,dernonstrating a sigiaificant difference compared with that at 0.5h and 8h. Expressions of TNF α mRNA from pancreas, liver and lung were decreased 50～80% in the treatment group, the psncreatie necrosis was also attenuated dramatically. Conclusiort. TNF α mRNA was detectable in pancreas,liver and lung tissues at the early stage of AHNP. SS-OP can significantly inhibit the expression of TNF α mRNA and attenuate the pancreatie necrosis.We feel that this may be an important mechanism of SS-OP in the treatment of AHNP,     

Effects of glycine and methylprednisolone on hemorrhagic shock in rats

Background Methylprednisolone (MP), a synthetic glucocorticosteroid, has been broadly studied in experiments on endotoxin-induced shock and septic shock. This study was designed to ascertain whether glycine and MP can protect against organ injury and death caused by hemorrhagic shock, and to elucidate the underlying mechanisms of these protective effects in rats.Method To establish a shock model, Wistar rats were bled to maintain mean arterial pressure at 30-50 mmHg for 1 hour and subsequently resuscitated with the shed blood and normal saline. Just prior to resuscitation, the rats were randomly assigned to four groups: sham group (operation performed without inducing shock), shock group, shock+glycine group (glycine injected at the beginning of resuscitation) and shock+MP group (MP injected at the beginning of resuscitation).Results ① Seventy-two hours after resuscitation, the survival rate of rats from the shock group had decreased to 20%, while the survival rates of rats from the shock+glycine and shock+MP groups were 77.8% and 80%, respectively. The difference was significant (P<0.05). ② Eighteen hours after resuscitation, pathological alterations in the organs of the rats were apparent. In rats from the shock group, edema, interstitial leukocyte infiltration, and cellular degeneration occurred in the liver, lungs, kidneys, and heart. Glycine and MP reduced these pathological changes significantly. ③ Eighteen hours after resuscitation, the levels of creatine phosphokinase, transaminases, and creatine were elevated significantly in rats from the shock group, indicating injury to the heart, liver, and kidneys, while these levels were elevated only slightly in the shock+glycine and shock+MP groups. The differences were significant (P<0.01). ④ There were significant increases in intracellular calcium and production of tumor necrosis factor (TNF-α) by isolated Kupffer cells stimulated by endotoxin after hemorrhagic shock. These changes were completely prevented by glycine and MP (P<0.01). Conclusion Glycine and MP reduce organ injury and mortality caused by hemorrhagic shock by preventing increase of intracellular calcium levels in Kupffer cell, suppressing Kupffer cell activation, decreasing the production of TNF-α by Kupffer cells, and blocking systemic inflammatory responses.     

Redox factor-1 may mediate the repair of multiple organ injuries after liver transplantation

AIM: To examine the relationship between multiple-organ injuries and expression of redox factor-1 (Ref-1) in the early period after liver transplantation. METHODS: A total of 150 adult male Wister rats were divided randomly into three groups: liver transplantation, sham surgery, and untreated control. After liver transplantation, animals were sacrificed at different time points. Hepatic and renal functions were analyzed by serology. Histology, apoptotic levels, and Ref-1 expression were examined by immunohistochemistry in the liver, kidneys, intestines, and lungs. RESULTS: Serum levels of alanine aminotransferase and aspartate aminotransferase peaked 6 h after liver transplantation and decreased appreciably after 12 h in the transplantation group, suggesting that the degree of liver injury in the early period after transplantation peaked at 6 h and then decreased. Pathological analyses showed that hepatic tissues were more severely injured in the transplantation group than in the sham and untreated groups. A considerable number of infiltrating inflammatory cells was observed around the portal vein in the transplantation group. Injuries to the kidneys, intestines, and lungs were milder after liver transplantation. Apoptotic levels increased after liver transplantation in all four organs examined. Ref-1 expression was higher in the transplantation group in the early period after liver transplantation than in the sham surgery and untreated control groups. CONCLUSION: Ref-1 expression induced by ischemia–reperfusion injury may have a critical role in repairing multiple-organ injuries after liver transplantation.     

Changes of Lipid Peroxides and Superoxide Dismutase in Visceral Tissues during Endotoxic Shock in Rats

As compared with the control,the malondialdehyde(MDA)levels in the cardi-ac,pulmonary,and intestinal tissues were elevated(P<0.05),and the superoxide dismu-tase activities in the cardiac,pulmonary,and intestinal tissues increased(P<0.05)at the30th min after endotoxic shock;the former in the cardiac,hepatic,renal,pulmonary,andintestinal tissues were elevated(P<0.05～0.001)and the latter in the cardiac,pulmona-ry,and intestinal tissues decreased in the 2nd h after shock;and the former were marke-dly elevated(P<0.01～0.001)and the latter severely decreased(P<<0.05～0.001)in allthe visceral tissues tested in the 4th h after shock.These data showed that lipid peroxid-ation mediated through oxygen-derived free radicals brought about tissue damage of theviscera listed above,which occurred earlier in the heart,the lungs and the intestines thanin the liver and the kidneys,and superoxide dismutase(SOD)activity was suppressed inthe late stage of endotoxic shock.     

ENDOTOXIN-INDUCED LIVER INJURY AND CHANGES IN THE LEVELS OF PLASMA TUMOR NECROSIS FACTOR-α AND INTERLEUKIN-6 IN RABBITS

ELISA was employed to detect changes in plasma TNF-α and IL-6 level in rabbits having endotoxin-induced generalized Shwartzman reaction. Liver injury was assessed by the increase of serum ALT and hepatic histopathologic changes. The results showed that plasma TNF-α and IL-6 levels increased remarkably 12h after intravenous injection of endotoxin twice at a 24h interval, and these changes corresponded with the degree of liver injury. Coinjection of dexamethasone and endotoxin partly prevented the elevation of TNF-α and IL-6 levels and reduced liver injury. These results indicated that TNF-α and IL-6 were involved in endotoxin-induced liver injury.     

Serum tumor necrosis factor (TNF) in the pathogenesis of clinical hepatic failure of HCV and/or HBV infection.

Serum TNF and IL-6 levels were measured in 48 patients with liver disease positive for anti-HCV only or concurrent HBV infection. High serum TNF levels were observed in patients with liver disease positive for anti-HCV and/or HBV infection (P < 0.001). Serum TNF levels varied with the severity of liver disease. Serum TNF levels of anti-HCV positive patients with hepatic failure were higher than those with CAH (P < 0.01). Serum TNF levels of patients infected with HCV or concurrent HBV were also significantly higher than those with HBV infection alone (P < 0.001). However, no difference in serum IL-6 levels was observed in either group of patients. Serum TNF in the deceased patients with hepatic failure induced by HBV and HCV infection was higher than in those who survived (P < 0.05), and it also seemed significantly different in patients with and without multiple organ failure (P < 0.05). In vitro, HSS showed marked inhibitory activity on TNF production from PBM induced by endotoxin, but had no significant effect on the TNF cytotoxicity of L929 cells. It seems that high serum TNF level is an important mediator in the pathogenesis of liver necrosis and failure of microcirculation in HCV and/or HBV infection. These observations favor the attempt to treat hepatic failure with HSS or anti-TNF. Encouraging results were achieved using HSS in the treatment of subacute liver necrosis in our institute.(ABSTRACT TRUNCATED AT 250 WORDS)

     

Endotoxin-induced liver injury and changes in the levels of plasma tumor necrosis factor-alpha and interleukin-6 in rabbits.

ELISA was employed to detect changes in plasma TNF-alpha and IL-6 level in rabbits having endotoxin-induced generalized Shwartzman reaction. Liver injury was assessed by the increase of serum ALT and hepatic histopathologic changes. The results showed that plasma TNF-alpha and IL-6 levels increased remarkably 12h after intravenous injection of endotoxin twice at a 24h interval, and these changes corresponded with the degree of liver injury. Coinjection of dexamethasone and endotoxin partly prevented the elevation of TNF-alpha and IL-6 levels and reduced liver injury. These results indicated that TNF-alpha and IL-6 were involved in endotoxin-induced liver injury.

     

EFFECT OF HEMORRHAGIC SHOCK ON ENDOTOXININDUCED TNF PRODUCTION AND ITS MOLECULAR MECHANISM IN RATS

INTRODUCTIONAlthoughmanyinvestigatorshavereportedthatonlymildendotoxemiaoccurredinhumansandanimalsafterhemorrhagicshock(1~3),bacterialendotoxinhasbeenshowntoplayanimportantroleinthedevelop-mentofshock,becauseanti-endotoxintreatmentcouldprotectshock-animalsfromdeath(2,4,5).Itremainsobscurewhethertheroleofendogenousendotoxininhemorrhagicshockisrelatedtoincreasedsensitivitytoendotoxin.Cytokines,suchastumornecrosisfactor(TNF)havebeenconsideredtobetheimportantmedi-atorsforregulationofendotoxi…     

Distribution of endotoxins in tissues and circulation and its effects following hemorrhagic shock

OBJECTIVE: To systemically investigate 1) distribution of endogenous endotoxin (ET) in tissues and circulation; 2) its relationship with shock duration and organ damage; and 3) its possible mechanism after hemorrhagic shock. METHODS: To further elucidate the intrinsic relationship between endogenous endotoxin translocation and hemorrhagic shock, the present study systematically investigated the distribution of endogenous ET into the liver, lungs, kidneys and circulation, and the relationship between ET levels and the corresponding organ dysfunction with limulus amebocyte lysate (LAL) chromogenic assay following hemorrhagic shock in rats. RESULTS: It was found that ET levels in hepatic homogenate markedly increased (P = 0.09) 1.5 hours following shock compared with that in the sham group. After resuscitation, ET levels in hepatic, pulmonary and renal tissues were all significantly elevated. The levels kept increasing with the prolonged experimental time, and reached as high as 3.88 +/- 0.95 EU (endotoxin unit)/g in the livers, 2.53 +/- 1.46 EU/g in the lungs and 2.51 +/- 0.89 EU/g in the kidneys 12 hours after shock. ET levels in plasma reached a peak of 1.13 +/- 0.42 EU/ml at 1 hour following resuscitation, then rapidly decreased to the sham levels 3 hours following resuscitation. There was a close relationship between endotoxin translocation and shock duration. Correlation analysis further indicated that the changes in glutamic-pyruvic transaminase (GPT), blood urea nitrogen (BUN) in plasma and angiotensin I-converting exzyme (ACE) in pulmonary homogenate were significantly and positively correlated with the ET levels in the liver, kidneys and lungs after hemorrhagic shock. CONCLUSIONS: Hemorrhagic shock can induce obvious endogenous ET translocation, which is closely related to the shock duration. Although only transient endotoxemia occurs after hemorrhagic shock, ET can massively accumulate in tissues (liver, lungs and kidneys), and may play an important role in the development of shock.     

Expression of TNF mRNA in the internal organs after severe burn injury in rats

ＥｘｐｒｅｓｓｉｏｎｏｆＴＮＦｍＲＮＡｉｎｔｈｅｉｎｔｅｒｎａｌｏｒｇａｎｓａｆｔｅｒｓｅｖｅｒｅｂｕｒｎｉｎｊｕｒｙｉｎｒａｔｓＹｕａｎＪｉａｎｃｈｅｎｇ；（袁建成）；ＸｉａｏＧｕａｎｇｘｉａ；（肖光夏）；ＺｈｏｕＬｉｘｉｎｇ；（周立新）；Ｑｉｎｇ...     

免疫吸附清除循环TNF对组织中TNF和IL-8水平的影响

用新西兰白兔建立内毒素休克模型，以亲和免疫吸附清除循环中TNF，观察对动物心、肝、肺、肾组织中TNF和IL－8水平的影响。结果表明，能有效清除循环中TNF，并使组织中TNF和IL－8水平明显降低。     

TNF在动物多器官衰竭中的变化及意义

在动物多器官衰竭模型上动态观察肿瘤坏死因子变化，探讨ＴＮＦ与ＭＯＦ的内在联系。４０只家兔分为３组：Ａ组２０只，Ｂ，Ｃ组各１０只。Ａ组用失血性休克，内毒素复合因素复制出ＭＯＦ模型；Ｂ组仅给内毒素；Ｃ组仅施失血性休克。衰竭器官≥２个者诊断为ＭＯＦ，ＴＮＦ测定用生物法。     

Role of tumor necrosis factor-alpha and interleukin-1beta on lung dysfunction following hemorrhagic shock in rats.

BACKGROUND: Hemorrhagic shock occasionally causes a fatal outcome following an outbreak of lung dysfunction, but the precise mechanism has not been clearly elucidated. Several studies have indicated that hemorrhagic shock causes a delayed vascular inflammatory decompensation and leads to inflammation-related organ dysfunction. Tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta are known as major proinflammatory cytokines that play an important role in excessive autolytic inflammation, finally inducing organ dysfunctions. In this study, the role of TNF-alpha and IL-1beta on lung dysfunction following hemorrhagic shock was examined by using FR167653, a potent inhibitor of TNF-alpha and IL-1beta production that acts by suppressing p38 mitogen-activated protein kinase (MAPK). MATERIAL/METHODS: Hemorrhagic shock was induced in anesthetized male rats by bleeding via a common carotid catheter for 20 minutes to 25% of total body blood volume without fluid resuscitation. Mean blood pressure, heart rate and arterial blood gas components were recorded up to 5 hours after the bleeding. The levels of TNF-alpha, IL-1beta and lactic dehydrogenase (LDH)-3 isozyme were measured in the serum of pulmonary venous blood. The lung tissue was excised for the assay of mRNA and for histopathological study. RESULTS: The expressions of mRNA for TNF-alpha and IL-1beta in the lung tissue and the concentrations of both cytokines in pulmonary serum increased after a hemorrhage. Inflammation-related injuries and function deterioration were observed in the lung following hemorrhagic shock. These hemorrhagic changes were inhibited by pretreatment with FR167653. CONCLUSIONS: TNF-alpha and IL-1beta play a key role in the development of inflammation-related lung dysfunction following hemorrhagic shock. Our model should be useful to explain the pathogenesis of lung dysfunction following hemorrhagic shock.     

失血性休克时脾脏TNF和IL-6生成的变化及氧合液复苏对其逆转作用

目的 探讨失血性休克对脾脏细胞因子生成的影响及氧合液复苏的调节作用。方法 取失血性休克家犬动静脉血及脾组织匀浆放免法测定肿瘤坏死因子（TNF）和白细胞介素-6（IL-6）水平。结果 休克后动脉及脾静脉血浆中TNF和IL-6水平明显降低，休克复苏后动静脉血浆中TNF和IL-6水平均明显升高，休克脾组织中TNF和IL-6水平明显增高，休克复苏后脾组织中TNF和IL-6水平进一步增高。结论 失血性休克可影响TNF和IL-6的生成和释放，氧合液复苏能较好的恢复其作用。     

肝癌患者血清IL—10，IL—12和TNFα水平的变化

OBJECTIVE: To study the relationship and clinical significance between level change of serum Interleukin 10 (IL-10), interleukin 12 (IL-12), tumor necrosis factor-alpha (TNF alpha) and primary hepatic carcinoma (PHC). METHODS: Serum IL-10, IL-12, TNF alpha in 40 patients with PHC, 36 cases with hepatocirrhosis and 33 normal controls were measured with ELISA. RESULTS: The higher serum levels of IL-10, IL-12, TNF alpha were observed in patients with hepatocirrhosis than in controls (P < 0.01), although IL-12, TNF alpha in the serum of patients with PHC were in the proximity of that of controls (P > 0.05), the IL-10 level was significantly higher than that of control (P < 0.01), further more, IL-10 being opposed to IL-12, TNF alpha is higher in patients with tumor above 5.0 cm in diameter than below 5.0 cm, IL-12 and TNF alpha were declined and IL-10 increased with tumor enlargement. CONCLUSION: The immune dysfunction in patients with PHC may be related to level increase of IL-10. The level decrease of IL-12, TNF alpha and increase of IL-10 is helpful to clinical differential diagnoses between cirrhotic nodule and PHC, and this suggests cancerous tendency.     

内毒素吸附剂对失血性休克大鼠 NO，IL-6及 IL-12水平的影响

目的:探讨内毒素吸附剂对失血性休克大鼠一氧化氮(NO),白细胞介素-6(IL-)及白细胞介素-2(IL-2)水平的影响。方法:选取成年SD大鼠50只,随机分为正常对照组10只,模型组和药物组各20只。模型组和药物组建立失血性休克大鼠模型,药物组造模前30 min采用内毒素吸附剂进行灌胃。复苏后1 h、2 h、4 h、8 h、12 h检测所有大鼠血清NO、IL-6、IL-2及内毒素水平。结果:模型组和药物组血清NO、IL-、IL-2及内毒素水平均高于正常对照组,且模型组高于药物组,差异具有统计学意义(P<0.05)。结论:内毒素吸附剂可明显降低失血性休克模型大鼠血清NO、IL-、IL-2及内毒素水平,对减轻炎性反应、减少组织损伤有显著作用。